Two Dimensional Gel Electrophoresis in Cancer Proteomics
نویسندگان
چکیده
Two-dimensional electrophoresis (2-DE) is a powerful and widely used method for the analysis of complex protein mixtures extracted from cells, tissues, or other biological samples. This technique sort’s protein according to two independent properties in two discrete steps: the first-dimension step, isoelectric focusing (IEF), separates proteins according to their isoelectric points (pI); the second-dimension step, SDS-polyacrylamide gel electrophoresis (SDS-PAGE), separates proteins according to their molecular weights (Mr, relative molecular weight). Each spot on the resulting two-dimensional array corresponds to a single protein species in the sample. Thousands of proteins can thus be separated, and information such as the protein pI, the apparent molecular weight, and the amount of each protein obtained. The separation of proteins by 2-DE dates back to the 1950s. The first 2-DE technique was developed by Smithies and Poulik in 1956 and O'Farrell, 1975 and Klose, 1975 significantly modified this method to elucidate protein profile. In the original technique, the first-dimension separation was performed in carrier ampholyte-containing polyacrylamide gels cast in narrow tubes.
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